Dendroctonus valens is an invasive pest in coniferous forests of northern China. It was suspected of being responsible for the death of more than three million Pinus tabuliformis trees. The present study sought to identify the ophiostomatoid fungi associated with D. valens in northern China and understand the possible role of these fungi in the pine decline. On the basis of morphology, physiology, mating compatibility and phylogenetic analyses of multiple DNA sequences, seven species of ophiostomatoid fungi were isolated from and around D. valens galleries: Leptographium alethinum, Grosmannia koreana (teleomorph of L. koreanum), L. procerum, L. sinoprocerum, L. truncatum, Pesotum aureum and P. pini. All have been recorded for the first time in China. Among them, the occurrence of the dominant species L. procerum is positively linked to attack intensities of D. valens. The pathogenicity of four species (L. koreanum, L. procerum, L. sinoprocerum and L. truncatum) was tested on mature P. tabuliformis trees by stem inoculation. All inoculated strains caused significant necrotic lesions on the inner bark. However, L. koreanum and L. truncatum induced more extensive lesions than L. procerum and L. sinoprocerum. Their association with D. valens and the P. tabuliformis decline is discussed.
Pine wilt disease (PWD) caused by pine wood nematode (PWN), Bursaphelenchus xylophilus, is the most destructive diseases of pine and poses a threat of serious economic losses worldwide. Although several of the mechanisms involved in disease progression have been discovered, the molecular response of Pinus massoniana to PWN infection has not been explored. We constructed four subtractive suppression hybridization cDNA libraries by taking time-course samples from PWN-inoculated Masson pine trees. One-hundred forty-four significantly differentially expressed sequence tags (ESTs) were identified, and 124 high-quality sequences with transcriptional features were selected for gene ontology (GO) and individual gene analyses. There were marked differences in the types of transcripts, as well as in the timing and levels of transcript expression in the pine trees following PWN inoculation. Genes involved in signal transduction, transcription and translation and secondary metabolism were highly expressed after 24 h and 72 h, while stress response genes were highly expressed only after 72 h. Certain transcripts responding to PWN infection were discriminative; pathogenesis and cell wall-related genes were more abundant, while detoxification or redox process-related genes were less abundant. This study provides new insights into the molecular mechanisms that control the biochemical and physiological responses of pine trees to PWN infection, particularly during the initial stage of infection.
The third-stage dispersal juvenile (JIII) is the stage for survival and dispersal in the winter of the pine wood nematode, Bursaphelenchus xylophilus. Through investigations at different temperatures, we found two kinds of growth-arrested development, including the adult longevity extension and JIII formation induced by low temperature. They showed similar characters: densely packed lipid droplets and extended longevity. We considered that there were four stages in the formation of growth-arrested stages: induction, growth-arrested pathway, growth-arrested development and cold-tolerance duration. Moreover, at 4 • C there were significant changes in secondary metabolites, which may be related to signal communication and metabolism associated with the formation of growth-arrested stages. The results suggested that low temperature was necessary for the dispersal of pine wood nematode and influenced distribution and intensity of pine wilt.
During a study of ophiostomatoid fungi associated with the invasive pest Dendroctonus valens in the Pinus tabuliformis ecosystem in northern China, a multigenic (ITS2-LSU, beta-tubulin and EF1-alpha) phylogenetic analysis and examination of morphological features revealed in addition to Leptographium procerum the occurrence of an undescribed species. The new species, Leptographium sinoprocerum, belongs to the L. procerum-L. profanum clade. Both L. procerum and L. sinoprocerum are similar to each other and occur sympatrically in the ecosystem studied. Nevertheless L. sinoprocerum can be distinguished from L. procerum by shorter conidiophore stipes arising from both submerged and aerial hyphae, slightly more oblong conidia, a granular ornamentation on the submerged hyphae and dark olivaceous colonies on MEA.
During a study of ophiostomatoid fungi associated with the invasive pest Dendroctonus valens in the Pinus tabuliformis ecosystem in northern China, a multigenic (ITS2-LSU, beta-tubulin and EF1-alpha) phylogenetic analysis and examination of morphological features revealed in addition to Leptographium procerum the occurrence of an undescribed species. The new species, Leptographium sinoprocerum, belongs to the L. procerum-L. profanum clade. Both L. procerum and L. sinoprocerum are similar to each other and occur sympatrically in the ecosystem studied. Nevertheless L. sinoprocerum can be distinguished from L. procerum by shorter conidiophore stipes arising from both submerged and aerial hyphae, slightly more oblong conidia, a granular ornamentation on the submerged hyphae and dark olivaceous colonies on MEA.
Pine wilt disease is a devastating forest disease worldwide caused by the pinewood nematode (PWN; Bursaphelenchus xylophilus). Esteya vermicola is widely recognized as a promising bio‐control agent effective against PWN. During a survey of associated microorganisms of Pinus yunnanensis in Southwest China, a novel isolate of E. vermicola was obtained. This isolate produced two types of conidia. One type of conidia was hyaline, unicellular, bacilloid, and non‐adhesive. The second type of conidia is solitary, hyaline, unicellular asymmetric elliptic or lunate, concave inward, ending slightly apiculate, adhesive on the concave surface, and contained an ovoid endospore‐like structure. Only the lunate conidia showed infection activity against PWN. This novel isolate produced a high proportion of infective lunate conidia and exhibited high adhesion and infection activity against PWN by adhering to the nematode within 48 h and killing 74.5% of the tested PWN individuals within 4 days. Given that the isolate was obtained from P. yunnanensis, it may be well adapted to the endogenous environment of pine trees. The isolate shows potential as a bio‐control agent against a plant‐parasitic nematode in the field.
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